Abstract
For plasmid-mediated gene therapy applications, a major limitation to scale up from rodents to large animals is the low expression level of injected plasmid DNA. The electroporation technique, which results in the passage of foreign material through the cell membrane, is one method that has been shown to be effective at improving local plasmid uptake and consequently, expression levels. Previous studies have determined that optimized electroporation parameters (such as electric field intensity, number of pulses, lag time between plasmid injections and electroporations, and optimal plasmid formulation conditions) are dependent on the target muscle type and individual species. Here, we provide a detailed protocol to optimize conditions for the successful intramuscular electroporation of plasmid DNA to swine, a large animal model. Our results suggest that the technique is safe and effective for veterinary applications. Furthermore, these results provide evidence for the feasibility of upcoming human applications.
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References
van Gaal, E.V., Hennink, W.E., Crommelin, D.J., Mastrobattista, E. (2006) Plasmid engineering for controlled and sustained gene expression for nonviral gene therapy. Pharm Res 23,1053–1074.
Draghia-Akli, R., Khan, A.S., Cummings, K.K., Parghi, D., Carpenter, R.H., Brown, P.A. (2002) Electrical enhancement of formulated plasmid delivery in animals. Tech Cancer Res Treat 1, 365–371.
Miyazaki, S., Miyazaki, J. (2008) In vivo DNA electrotransfer into muscle. Dev Growth Differ 50, 479–483.
Khan, A.S., Smith, L.C., Abruzzese, R.V., Cummings, K.K., Pope, M.A., Brown, P.A., Draghia-Akli, R. (2003) Optimization of electroporation parameters for the intramuscular delivery of plasmids in pigs. DNA Cell Biol 22, 807–814.
Khan, A.S., Pope, M.A., Draghia-Akli, R. (2005) Highly efficient constant-current electroporation increases in vivo plasmid expression. DNA & Cell Biology 24, 810–818.
Draghia-Akli, R., Khan, A.S., Brown, P.A., Pope, M.A., Wu, L., Hirao, L., Weiner, D.B. (2008) Parameters for DNA vaccination using adaptive constant-current electroporation in mouse and pig models. Vaccine 26, 5230–5237.
Schertzer, J.D., Plant, D.R., Lynch, G.S. (2006) Optimizing plasmid-based gene transfer for investigating skeletal muscle structure and function. Mol Ther 13, 795–803.
Tone, C.M., Cardoza, D.M., Carpenter, R.H., Draghia-Akli, R. (2004) Long-term effects of plasmid-mediated growth hormone releasing hormone in dogs. Cancer Gene Ther 11, 389–396.
Draghia-Akli, R., Cummings, K.K., Khan, A.S., Brown, P.A., Carpenter, R.H. (2003) Effects of plasmid-mediated growth hormone releasing hormone supplementation in young healthy Beagle dogs. J Anim Sci 81, 2301–2310.
Bodles-Brakhop, A.M., Brown, P.A., Pope, M.A., Draghia-Akli, R. (2008) Double-blinded, placebo-controlled plasmid GHRH trial for cancer-associated anemia in dogs. Mol Ther 16, 862–870.
Brown, P.A., Bodles-Brakhop, A.M., Pope, M.A., Draghia-Akli, R. (2009) Gene therapy by electroporation for the treatment of chronic renal failure in companion animals. BMC Biotechnol 9, 4.
Brown, P.A., Bodles-Brakhop, A., Draghia-Akli, R. (2008) Plasmid growth hormone releasing hormone therapy in healthy and laminitis-afflicted horses-evaluation and pilot study. J Gene Med 10, 564–574.
Draghia-Akli, R., Ellis, K.M., Hill, L.A., Malone, P.B., Fiorotto, M.L. (2003) High-efficiency growth hormone releasing hormone plasmid vector administration into skeletal muscle mediated by electroporation in pigs. FASEB J 17, 526–528.
Person, R., Bodles-Brakhop, A.M., Pope, M.A., Brown, P.A., Khan, A.S., Draghia-Akli, R. (2008) Growth hormone-releasing hormone plasmid treatment by electroporation decreases offspring mortality over three pregnancies. Mol Ther 16, 1891–1897.
Brown, P.A., Davis, W.C., Draghia-Akli, R. (2004) Immune enhancing effects of growth hormone releasing hormone delivered by plasmid injection and electroporation. Mol Ther 10, 644–651.
Brown, P.A., Bodles-Brakhop, A.M., Draghia-Akli, R. (2008) Effects of plasmid growth hormone releasing hormone treatment during heat stress. DNA & Cell Biology 27, 629–635.
Draghia-Akli, R., Fiorotto, M.L., Hill, L.A., Malone, P.B., Deaver, D.R., Schwartz, R.J. (1999) Myogenic expression of an injectable protease-resistant growth hormone-releasing hormone augments long-term growth in pigs. Nat Biotechnol 17, 1179–1183.
Khan, A.S., Fiorotto, M.L., Cummings, K.K., Pope, M.A., Brown, P.A., Draghia-Akli, R. (2003) Maternal GHRH plasmid administration changes pituitary cell lineage and improves progeny growth of pigs. Am J Physiol Endocrinol Metab 285, E224–E231.
Acknowledgments
The authors would like to thank Dr. Patricia Brown for her help as an attending veterinarian to oversee the animal anesthesia and perform the electroporation animal procedures. This work was supported by Inovio Pharmaceuticals (Blue Bell, PA).
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Bodles-Brakhop, A.M., Draghia-Akli, R., Broderick, K., Khan, A.S. (2011). Electroporation of Plasmid DNA to Swine Muscle. In: Duan, D. (eds) Muscle Gene Therapy. Methods in Molecular Biology, vol 709. Humana Press. https://doi.org/10.1007/978-1-61737-982-6_16
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DOI: https://doi.org/10.1007/978-1-61737-982-6_16
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Publisher Name: Humana Press
Print ISBN: 978-1-61737-981-9
Online ISBN: 978-1-61737-982-6
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